Abstract
Aldehyde oxidase and xanthine oxidoreductase are part of the molybdo-flavo family of enzymes. They have become increasingly relevant in drug discovery as medicinal chemistry strategies evolve to reduce cytochrome P450 (CYP) related drug metabolism. Consequently, there is a need for a differentiation strategy for test compounds to be characterized from those of CYP and molybdenum-containing hydroxylases. Herein, general procedures to identify activity and differentiate the fraction metabolized by aldehyde oxidase and xanthine oxidoreductase from that of the family of CYP enzymes are described. Additionally, identifying structural features that lend themselves to aldehyde oxidase and xanthine oxidoreductase metabolism, tissue fractions appropriate to measure activities of these enzymes, specific cofactors to add or omit, specific inhibitors, kinetic idiosyncrasies, and confirmatory studies are detailed.
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References
Beedham C (2001) Molybdenum hydroxylases, in enzyme systems that metabolize drugs and other xenobiotics. John Wiley & Sons, Ltd, Chichester, West Sussex, UK, pp 147–187
Kitamura S, Sugihara K, Ohta S (2006) Drug-metabolizing ability of molybdenum hydroxylases. Drug Metab Pharmacokinet 21:83–98
Garattini E, Fratelli M, Terao M (2008) Mammalian aldehyde oxidase: genetics, evolution and biochemistry. Cell Mol Life Sci 65:1019–1048
Pryde DC, Dalvie D, Hu Q, Jones P, Obach RS, Tran TD (2010) Aldehyde oxidase: an enzyme of emerging importance in drug discovery. J Med Chem 53:8441–8460
Garattini E, Terao M (2012) The role of aldehyde oxidase in drug metabolism. Expert Opin Drug Metab Toxicol 8:487–503
Battelli MG, Polito L, Bortolotti M, Bolognesi A (2016) Xanthine oxidoreductase in drug metabolism: beyond a role as a detoxifying enzyme. Curr Med Chem 23:4027–4036
Diamond S, Boer J, Maduskuie TP Jr, Falahatpisheh N, Li Y, Yeleswaram S (2010) Species-specific metabolism of SGX523 by aldehyde oxidase and the toxicological implications. Drug Metab Dispos 38:1277–1285
Akabane T, Tanaka K, Irie M, Terashita S, Teramura T (2011) Case report of extensive metabolism by aldehyde oxidase in humans: pharmacokinetics and metabolite profile of FK3453 in rats, dogs, and humans. Xenobiotica 41:372–384
Jensen KG, Jacobsen AM, Bundgaard C, Nilausen DO, Thale Z, Chandresena G, Jørgensen M (2017) Lack of exposure in a first-in-man study due to aldehyde oxidase metabolism: investigated by use of 14C-microdose, humanized mice, monkey pharmacokinetics, and in vitro methods. Drug Metab Dispos 45:68–75
Pryde DC, Tran TD, Jones P, Duckworth J, Howard M, Gardner I, Hyland R, Webster R, Wenham T, Bagal S, Omoto K, Schneider RP, Lin J (2012) Medicinal chemistry approaches to avoid aldehyde oxidase metabolism. Bioorg Med Chem Lett 22:2856–2860
Terao M, Romão MJ, Leimkühler S, Bolis M, Fratelli M, Coelho C, Santos-Silva T, Garattini E (2016) Structure and function of mammalian aldehyde oxidases. Arch Toxicol 90:753–780
Zientek MA, Youdim K (2015) Reaction phenotyping: advances in the experimental strategies used to characterize the contribution of drug-metabolizing enzymes. Drug Metab Dispos 43:163–181
Barr JT, Jones JP, Joswig-Jones CA, Rock DA (2013) Absolute quantification of aldehyde oxidase protein in human liver using liquid chromatography-tandem mass spectrometry. Mol Pharm 10:3842–3849
Fu C, Di L, Han X, Soderstrom C, Synder M, Troutman MD, Obach RS, Zhang H (2013) Aldehyde oxidase 1 (AOX1) in human liver cytosols: quantitative characterization of AOX1 expression levels and activity relationship. Drug Metab Dispos 41:1797–1804
Feng L, Zhao N, Yao X, Sun X, Du L, Diao X, Li S, Li Y (2007) Histidine-tryptophan-ketoglutarate solution vs. University of Wisconsin solution for liver transplantation: a systematic review. Liver Transpl 13:1125–1136
Barr JT, Choughule KV, Nepal S, Wong T, Chaudhry AS, Joswig-Jones CA, Zientek M, Strom SC, Schuetz EG, Thummel KE, Jones JP (2014) Why do most human liver cytosol preparations lack xanthine oxidase activity? Drug Metab Dispos 42:695–699
Helmstetter S, Lyon KC, Yerino P, Hilgedick A, Hatfield NM, Ewy B, Woodworth Z, Stanley F, Buckley DB, Olgilvie BW (2017) The effects of organ preservation solution on aldehyde oxidase and xanthine oxidase activity in pooled human liver S9. ISSX 14th European Meeting
Zientek M, Jiang Y, Youdim K, Obach RS (2010) In vitro-in vivo correlation for intrinsic clearance for drugs metabolized by human aldehyde oxidase. Drug Metab Dispos 38:1322–1327
Hutzler JM, Yang YS, Albaugh D, Fullenwider CL, Schmenk J, Fisher MB (2012) Characterization of aldehyde oxidase enzyme activity in cryopreserved human hepatocytes. Drug Metab Dispos 40:267–275
Kanika CV, Barnaba C, Joswig-Jones CA, Jones JP (2014) In vitro metabolism of 6-mercaptopurine in human liver: insights into the role of molybdoflavoenzymes aldehyde oxidase, xanthine oxidase, and xanthine dehydrogenase. Drug Metab Dispos 42:1334–1340
Strelevitz TJ, Orozco CC, Obach RS (2012) Hydralazine as a selective probe inactivator of aldehyde oxidase in human hepatocytes: estimation of the contribution of aldehyde oxidase to metabolic clearance. Drug Metab Dispos 40:1441–1448
Linder CD, Renaud NA, Hutzler MJ (2009) Is 1-aminobenzotriazole an appropriate in vitro tool as a nonspecific cytochrome P450 inactivator? Drug Metab Dispos 37:10–13
Ortiz de Montellano PR (2018) 1-Aminobenzotriazole: a mechanism-based cytochrome P450 inhibitor and probe of cytochrome P450 biology. Med Chem (Los Angeles) 8:38–65
Dick RA (2018) Refinement of in vitro methods for identification of aldehyde oxidase substrates reveals metabolites of kinase inhibitors. Drug Metab Dispos 46:846–859
Armina A, Paragas EM, Joswig-Jones CA, Rodgers JT, Jones JP (2019) Time course of aldehyde oxidase and why it is nonlinear. Drug Metab Dispos 47:473–483
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Kozminski, K.D., Zientek, M.A. (2021). Differentiation of Cytochrome P450-Mediated from Non-CYP-Mediated Metabolism: Aldehyde Oxidase and Xanthine Oxidoreductase. In: Yan, Z., Caldwell, G.W. (eds) Cytochrome P450. Methods in Pharmacology and Toxicology. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1542-3_17
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DOI: https://doi.org/10.1007/978-1-0716-1542-3_17
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